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. 2020 Mar 3;8:127. doi: 10.3389/fcell.2020.00127

FIGURE 5.

FIGURE 5

Knockdown of IFT88 impairs cilium function and decreases expression of HO-1 in FSS-treated NP cells. (A) IFT88 Knockdown. NP cells were transfected with negative control siRNA (NC-siRNA) and three different IFT88-siRNA (#1, #2, #3). Then, cells were subjected to western blotting for expression of IFT88 (A). Quantitative data from three independent experiments (B). The #1 IFT88-siRNA was used for the following experiments in this study. (C,D) Qualification of cilium prevalence (C) and cilium length (D) in NP cells from three independent experiments. NP cells were transfected with negative control siRNA (NC-siRNA) and IFT88 siRNA. Then, cells were treated with 12 dyne/cm2 FSS for 2 h. (E) Fluorescence images of primary cilia. Left bar, 10 μm; right bar, 5 μm. White arrows represent primary cilia. (F,G) Western blotting. NP cells were transfected with NC-siRNA or IFT88-siRNA (#1) and subjected to 12 dyne/cm2 FSS for 2 h, followed by western blotting for expression of IFT88 and HO-1 (F). Quantification of data from three independent experiments (G). Results are expressed as mean ± standard deviation (s.d.). *P < 0.05, **P < 0.01, and ***P < 0.001 vs. NC or FSS + NC-siRNA group).